DOCSLIB.ORG

MEIOB Exhibits Single-Stranded DNA-Binding and Exonuclease Activities and Is Essential for Meiotic Recombination

Total Page：16

File Type：pdf, Size：1020Kb

Download full-text PDF Read full-text
MEIOB Exhibits Single-Stranded DNA-Binding and Exonuclease Activities and Is Essential for Meiotic Recombination ARTICLE Received 20 Sep 2013 | Accepted 16 Oct 2013 | Published 18 Nov 2013 DOI: 10.1038/ncomms3788 MEIOB exhibits single-stranded DNA-binding and exonuclease activities and is essential for meiotic recombination Mengcheng Luo1, Fang Yang1, N. Adrian Leu1, Jessica Landaiche2, Mary Ann Handel3, Ricardo Benavente4, Sophie La Salle2 & P. Jeremy Wang1 Meiotic recombination enables the reciprocal exchange of genetic material between parental homologous chromosomes, and ensures faithful chromosome segregation during meiosis in sexually reproducing organisms. This process relies on the complex interaction of DNA repair factors and many steps remain poorly understood in mammals. Here we report the identi- fication of MEIOB, a meiosis-specific protein, in a proteomics screen for novel meiotic chromatin-associated proteins in mice. MEIOB contains an OB domain with homology to one of the RPA1 OB folds. MEIOB binds to single-stranded DNA and exhibits 30–50 exonuclease activity. MEIOB forms a complex with RPA and with SPATA22, and these three proteins co-localize in foci that are associated with meiotic chromosomes. Strikingly, chromatin localization and stability of MEIOB depends on SPATA22 and vice versa. Meiob-null mouse mutants exhibit a failure in meiosis and sterility in both sexes. Our results suggest that MEIOB is required for meiotic recombination and chromosomal synapsis. 1 Center for Animal Transgenesis and Germ Cell Research, Department of Animal Biology, University of Pennsylvania School of Veterinary Medicine, 3800 Spruce Street, 200E Old Vet, Philadelphia, Pennsylvania 19104, USA. 2 Department of Biochemistry, Chicago College of Osteopathic Medicine, Midwestern University, Downers Grove, Illinois 60515, USA. 3 The Jackson Laboratory, 600 Main Street, Bar Harbor, Maine 04609, USA. 4 Department of Cell and Developmental Biology, Biocenter of the University of Wu¨rzburg, Am Hubland, Wu¨rzburg 97074, Germany. Correspondence and requests for materials should be addressed to P.J.W. (email: [email protected]). NATURE COMMUNICATIONS | 4:2788 | DOI: 10.1038/ncomms3788 | www.nature.com/naturecommunications 1 & 2013 Macmillan Publishers Limited. All rights reserved. ARTICLE NATURE COMMUNICATIONS | DOI: 10.1038/ncomms3788 eiosis, the hallmark of gametogenesis in sexually cells14, and from XXY* testes, which lack all germ cells and thus reproducing organisms, creates haploid germ cells from contain only somatic cells15. By subtracting these two data sets Mdiploid progenitors and ensures their genetic diversity from the P18 data set, with a stringent cutoff value of at least 3 through recombination. Physical links between homologous unique peptides in P18 but none in P6 and XXY*, we identified 51 chromosomes permit the formation of crossovers required for putative meiotic chromatin-associated proteins (Fig. 1d and allele recombination and also facilitate accurate chromosome Supplementary Table S1). segregation at the first meiotic cell division1,2. Meiotic Identification of previously known meiotic chromatin-asso- recombination is initiated at DNA double-strand breaks (DSBs) ciated proteins confirmed the validity of our approach: 13 of generated by SPO11 (ref. 3); these DSBs become resected from 51 proteins identified are known to localize to meiotic the 50-end, yielding extended 30-single-stranded DNA (ssDNA) chromatin (SYCP1, SYCP2, SYCP3, SYCE2, SMC1B, HOR- overhangs4. Replication protein A (RPA) coats ssDNA, forming MAD1, H2AX, MDC1, TERF1, TDP1, PIWIL1, PIWIL2 and nucleoprotein filaments that prevent degradation and secondary YBX2; Supplementary Table S1), including all three synaptone- structure formation. Subsequent invasion of the 30-single strand mal complex proteins. Furthermore, the genes encoding 19 into the duplex of the homologue is mediated by DMC1 and proteins identified in our screen had been subjected to targeted RAD51. Strand invasion causes the displacement of one strand deletion in mice: 11 mutants exhibited meiotic arrest, 2 showed of the homologue, resulting in D-loop formation5. The invading post-meiotic spermiogenic arrest, 4 displayed embryonic lethality 30-end primes new DNA synthesis, such that D-loop extension (with an unknown role in meiosis) and 2 were dispensable for towards the second end allows for the capture of the 30-ssDNA fertility. The identification of these known meiosis factors end of the other break (second end). End ligation leads to the validates our biochemical/proteomic approach and suggests formation of double Holliday junctions. Although early steps of that the other 32 proteins identified in our screen are likely to meiotic recombination (DSB formation, end resection, strand be novel meiotic chromatin-associated proteins (Supplementary invasion and D-loop formation) have been relatively well Table S1). characterized, the intermediate steps of this process, such as second-end capture, are poorly defined. MEIOB is a novel meiosis-specific protein. To test whether the RPA is a conserved ssDNA-binding heterotrimer complex of newly identified putative meiotic chromatin-associated proteins RPA1, RPA2 and RPA3 (ref. 6). The four oligonucleotide-binding indeed localize to meiotic chromatin and function in meiosis, we (OB) folds of RPA1 mediate ssDNA binding of the RPA focused on one of these uncharacterized proteins—MEIOB. complex7. RPA is essential for many aspects of DNA MEIOB (470 amino acids (aa)) is predicted to contain an OB fold metabolism, notably DNA replication, DNA repair and meiotic (aa 167–272) with limited homology to the ssDNA-binding recombination. During meiotic recombination, RPA binds to domain-B OB domain of RPA1 (Supplementary Fig. S1). MEIOB various ssDNA intermediates, including resected 30-ssDNA and homologues are present in mammals, chicken, fugu fish, Droso- the D-loop. Studies of mutant alleles of yeast RPA1 have phila and Arabidopsis. Western blot analyses using specific anti- demonstrated that RPA is required for meiotic recombination8. bodies raised against MEIOB confirmed its association with RPA is indispensible for mouse development; a homozygous chromatin; MEIOB was also detectable in the cytoplasmic and point mutation in Rpa1 causes early embryonic lethality9. soluble nuclear fractions of the testis (Fig. 1e). Although abun- Here we report the identification of meiosis-specific with OB dantly expressed in the testis, MEIOB was not detectable in domains (MEIOB), a meiosis-specific protein with homology to somatic tissues when examined by western blotting (Fig. 1f). one OB fold of RPA1, in a systematic proteomics screen for MEIOB localized predominantly to the nuclei of spermatocytes in meiotic chromatin-associated proteins. We find that mouse the testis (Supplementary Fig. S2). Therefore, MEIOB is a novel MEIOB has evolved to fulfill an essential function in meiosis. meiotic chromatin-associated protein. Results MEIOB co-localizes with RPA in foci on meiotic chromosomes. Proteomics screen for meiotic chromatin-associated proteins. Immunostaining of spread nuclei of spermatocytes showed that To systematically identify proteins that are associated with MEIOB formed discrete foci on chromosomes, foci that exhibited meiotic chromosomes, we isolated chromatin from postnatal day a dynamic distribution pattern during meiosis (Fig. 2a–j). MEIOB 18 (P18) testes, which lack post-meiotic germ cells and are thus foci were absent at the early leptotene stage (Fig. 2a), became first highly enriched for meiotic germ cells. Our adaptation of a detectable on late leptotene chromosomes (Fig. 2b), formed arrays chromatin purification protocol for testis samples yielded highly along synaptonemal complexes in zygotene and early pachytene pure chromatin (Fig. 1a)10,11. SYCP2, a synaptonemal complex spermatocytes (Fig. 2c,d), and disappeared in late pachytene protein associated with meiotic chromatin12, was present in the spermatocytes (Fig. 2e). At a range of 100–200 per cell, MEIOB chromatin fraction, whereas the cytoplasmic germ cell-specific foci peaked at the zygotene stage (Fig. 2k). Notably, MEIOB foci protein TEX19 was not detectable in the chromatin fraction by were formed along the partially synapsed sex chromosomes at the western blot analysis (Fig. 1b)13. To improve detection of proteins pachytene stage (Fig. 2d). In addition, MEIOB formed foci on with low abundance, we separated chromatin-associated proteins meiotic chromosomes in fetal oocytes (Supplementary Fig. S3), from P18 testes by SDS–polyacrylamide gel electrophoresis implying functional relevance of MEIOB for meiosis in both (PAGE) and analysed ten different size fractions by tandem sexes. MEIOB was absent from the adult ovary (Fig. 1f), in which mass spectrometry (MS/MS; Fig. 1c). A total of 1,300 proteins oocytes are at the diplotene stage. No focal staining was observed with at least one unique peptide were identified. in Meiob À / À spermatocytes with the antibody used for these In addition to meiotic cells, P18 testes contain mitotic germ analyses, demonstrating that the antibody is specific for MEIOB cells and somatic cells (Fig. 1d). To identify proteins only and does not crossreact with RPA1 (Supplementary Fig. S4a). associated with meiotic chromatin, we applied a subtractive We next evaluated the potential interaction of MEIOB with approach, excluding chromatin-associated proteins present in other meiotic recombination-related proteins and found that mitotic cells. Using the same biochemical/proteomics protocol, MEIOB co-localized with RPA, which forms foci on prophase I we identified chromatin-associated proteins from postnatal
Load more

Recommended publications
  • Molecular and Physiological Basis for Hair Loss in Near Naked Hairless and Oak Ridge Rhino-Like Mouse Models: Tracking the Role of the Hairless Gene
    University of Tennessee, Knoxville TRACE: Tennessee Research and Creative Exchange Doctoral Dissertations Graduate School 5-2006 Molecular and Physiological Basis for Hair Loss in Near Naked Hairless and Oak Ridge Rhino-like Mouse Models: Tracking the Role of the Hairless Gene Yutao Liu University of Tennessee - Knoxville Follow this and additional works at: https://trace.tennessee.edu/utk_graddiss Part of the Life Sciences Commons Recommended Citation Liu, Yutao, "Molecular and Physiological Basis for Hair Loss in Near Naked Hairless and Oak Ridge Rhino- like Mouse Models: Tracking the Role of the Hairless Gene. " PhD diss., University of Tennessee, 2006. https://trace.tennessee.edu/utk_graddiss/1824 This Dissertation is brought to you for free and open access by the Graduate School at TRACE: Tennessee Research and Creative Exchange. It has been accepted for inclusion in Doctoral Dissertations by an authorized administrator of TRACE: Tennessee Research and Creative Exchange. For more information, please contact [email protected]. To the Graduate Council: I am submitting herewith a dissertation written by Yutao Liu entitled "Molecular and Physiological Basis for Hair Loss in Near Naked Hairless and Oak Ridge Rhino-like Mouse Models: Tracking the Role of the Hairless Gene." I have examined the final electronic copy of this dissertation for form and content and recommend that it be accepted in partial fulfillment of the requirements for the degree of Doctor of Philosophy, with a major in Life Sciences. Brynn H. Voy, Major Professor We have read this dissertation and recommend its acceptance: Naima Moustaid-Moussa, Yisong Wang, Rogert Hettich Accepted for the Council: Carolyn R.
    [Show full text]
  • A Computational Approach for Defining a Signature of Β-Cell Golgi Stress in Diabetes Mellitus
    Page 1 of 781 Diabetes A Computational Approach for Defining a Signature of β-Cell Golgi Stress in Diabetes Mellitus Robert N. Bone1,6,7, Olufunmilola Oyebamiji2, Sayali Talware2, Sharmila Selvaraj2, Preethi Krishnan3,6, Farooq Syed1,6,7, Huanmei Wu2, Carmella Evans-Molina 1,3,4,5,6,7,8* Departments of 1Pediatrics, 3Medicine, 4Anatomy, Cell Biology & Physiology, 5Biochemistry & Molecular Biology, the 6Center for Diabetes & Metabolic Diseases, and the 7Herman B. Wells Center for Pediatric Research, Indiana University School of Medicine, Indianapolis, IN 46202; 2Department of BioHealth Informatics, Indiana University-Purdue University Indianapolis, Indianapolis, IN, 46202; 8Roudebush VA Medical Center, Indianapolis, IN 46202. *Corresponding Author(s): Carmella Evans-Molina, MD, PhD ([email protected]) Indiana University School of Medicine, 635 Barnhill Drive, MS 2031A, Indianapolis, IN 46202, Telephone: (317) 274-4145, Fax (317) 274-4107 Running Title: Golgi Stress Response in Diabetes Word Count: 4358 Number of Figures: 6 Keywords: Golgi apparatus stress, Islets, β cell, Type 1 diabetes, Type 2 diabetes 1 Diabetes Publish Ahead of Print, published online August 20, 2020 Diabetes Page 2 of 781 ABSTRACT The Golgi apparatus (GA) is an important site of insulin processing and granule maturation, but whether GA organelle dysfunction and GA stress are present in the diabetic β-cell has not been tested. We utilized an informatics-based approach to develop a transcriptional signature of β-cell GA stress using existing RNA sequencing and microarray datasets generated using human islets from donors with diabetes and islets where type 1(T1D) and type 2 diabetes (T2D) had been modeled ex vivo. To narrow our results to GA-specific genes, we applied a filter set of 1,030 genes accepted as GA associated.
    [Show full text]
  • A Mutation in the Putative MLH3 Endonuclease Domain Confers a Defect in Both Mismatch Repair and Meiosis in Saccharomyces Cerevisiae
    Copyright Ó 2008 by the Genetics Society of America DOI: 10.1534/genetics.108.086645 A Mutation in the Putative MLH3 Endonuclease Domain Confers a Defect in Both Mismatch Repair and Meiosis in Saccharomyces cerevisiae K. T. Nishant, Aaron J. Plys and Eric Alani1 Department of Molecular Biology and Genetics, Cornell University, Ithaca, New York 14853-2703 Manuscript received January 2, 2008 Accepted for publication March 20, 2008 ABSTRACT Interference-dependent crossing over in yeast and mammalian meioses involves the mismatch repair protein homologs MSH4-MSH5 and MLH1-MLH3. The MLH3 protein contains a highly conserved metal- binding motif DQHA(X)2E(X)4E that is found in a subset of MLH proteins predicted to have endonuclease activities (Kadyrov et al. 2006). Mutations within this motif in human PMS2 and Saccharomyces cerevisiae PMS1 disrupted the endonuclease and mismatch repair activities of MLH1-PMS2 and MLH1-PMS1, re- spectively (Kadyrov et al. 2006, 2007; Erdeniz et al. 2007). As a first step in determining whether such an activity is required during meiosis, we made mutations in the MLH3 putative endonuclease domain motif (-D523N, -E529K) and found that single and double mutations conferred mlh3-null-like defects with respect to meiotic spore viability and crossing over. Yeast two-hybrid and chromatography analyses showed that the interaction between MLH1 and mlh3-D523N was maintained, suggesting that the mlh3-D523N mutation did not disrupt the stability of MLH3. The mlh3-D523N mutant also displayed a mutator phenotype in vegetative growth that was similar to mlh3D. Overexpression of this allele conferred a dominant-negative phenotype with respect to mismatch repair.
    [Show full text]
  • Research in Biology Using Computation Doi
    Journal of Bioinformatics and Genomics 2 (7) 2018 RESEARCH IN BIOLOGY USING COMPUTATION DOI: https://doi.org/10.18454/jbg.2018.2.7.1 Grishaeva T.M.* Department of Cytogenetics, Vavilov Institute of General Genetics, Russian Academy of Sciences, Moscow, Russian Federation * Correspodning author (grishaeva[at]vigg.ru) Received: 18.03.2018; Accepted: 05.04.2018; Published: 22.05.2018 COMPARATIVE CONSERVATION OF MEIOTIC PROTEINS IN DIFFERENT PHYLOGENETIC LINES OF EUKARYOTES Research article Abstract Motivation: Meiosis — a two-stage process of sex cell division — is served by several hundreds of proteins. A part of them went to eukaryotes from prokaryotes, others appeared in first eukaryotes, and some proteins appeared de novo in multicellular eukaryotes. We compared the conservation of proteins involved in various processes occurring in meiosis. Results: The conservations of five meiotic enzymes (MLH1, MRE11, MSH4, BRCA1, BRCA2) and three silencing markers (histone H2AX, SUMO1, ATR) were compared using a set of bioinformatics methods. Orthologs of these proteins from the proteomes of model species were compared, representing different lines of development of eukaryotes. Among the enzymes, the most conserved is MLH1, which provide correction of mismatch bases, and the least conserved are BRCA1 and BRCA2 repair enzymes which are present only in vertebrates. Among silencing proteins, histone H2AX is the most conserved one, playing the central part in the regulation of the transcription, in the repair and replication of DNA. The small protein SUMO1, which is involved in many cellular processes, is less conserved. ATR kinase in different species is similar only in the C-terminal part of the molecule.
    [Show full text]
  • Supplemental Data
    Supplementary Materials 1. Supplementary Methods 2. Supplementary Figures • Figure S1. RNA-seq measurement of kidney gene expression for known markers of acute kidney injury (Kim-1, Fgβ) and kidney fibrosis (Col1a1, Fn1). • Figure S2. Histological, biochemical and molecular characteristics of mouse models of kidney fibrosis and acute kidney injury. • Figure S3. RNA-seq vs. qRT-PCR correlation of gene expression measurement for the 11 selected candidates. • Figure S4. Histological characteristics of a normal vs. fibrotic human kidney. 3. Supplementary Tables • Table S1. List of genes identified by RNA-seq as significantly different from normal at least at one time-point in the FA nephropathy progression. • Table S2. List of peptides used for selected reaction monitoring (SRM) assays. • Table S3. List of primers used for mouse qRT-PCR. • Table S4. List of primers used for human qRT-PCR. Page 1 of 31 1. Supplementary Methods Animal studies Biospecimen collection: At the moment of sacrifice, blood was collected from the inferior vena cava under isoflurane anesthesia, and, following opening of the thoracic cavity to ensure that the animal is deceased, the kidneys were retrieved and sectioned in samples dedicated for histology and immunfluorescence (fixed in 10% neutral buffered formalin), protein and RNA analysis (flash-frozen in liquid nitrogen). Similarly, liver tissue sections from the left lateral lobe of the ANIT fed mice were fixed in neutral buffered formalin for histopathological processing, while other liver sections were flash-frozen in liquid nitrogen. Blood was collected from mice in heparinized tubes and plasma was separated following centrifugation at 7500 g for 5 minutes. Blood urea nitrogen (BUN) was measured using an InfinityUrea kit (Thermo Fisher Scientific, Wilmington, DE) and serum creatinine (SCr) was measured using a Creatinine Analyzer II (Beckman Coulter).
    [Show full text]
  • Human Muts Homologue MSH4 Physically Interacts with Von Hippel-Lindau Tumor Suppressor-Binding Protein 11
    [CANCER RESEARCH 63, 865–872, February 15, 2003] Human MutS Homologue MSH4 Physically Interacts with von Hippel-Lindau Tumor Suppressor-binding Protein 11 Chengtao Her,2 Xiling Wu, Michael D. Griswold, and Feng Zhou School of Molecular Biosciences and Center for Reproductive Biology, Washington State University, Pullman, Washington 99164-4660 [C. H., X. W., M. D. G.], and Bioscience Division, Los Alamos National Laboratory, Los Alamos, New Mexico 87545 [F. Z.] ABSTRACT human MMR genes are linked to the pathogenesis of hereditary nonpolyposis colorectal cancer (HNPCC) and sporadic tumors Increasing evidence indicated that the protein factors involved in DNA associated with microsatellite instability (1). Eukaryotic MutS mismatch repair (MMR) possess meiotic functions beyond the scope of DNA mismatch correction. The important roles of MMR components in homologous proteins MSH2, MSH3, and MSH6 are proteins meiotic processes have been highlighted by the recent identification of two known to participate in DNA MMR through the actions of their additional members of the mammalian MutS homologs, MSH4 and heterodimeric complexes consisting of either MSH2-MSH3 or MSH5. Mammalian MSH4 and MSH5 proteins form a heterodimeric MSH2-MSH6, in which the MSH2-MSH6 heterodimer recognizes complex and play an important role in the meiotic processes. As a step both single-base mismatches and small loops formed by insertions forward to the understanding of the molecular mechanisms underlying or deletions in the DNA, whereas the MSH2-MSH3 heterodimer the roles of these two mammalian MutS homologues, here we have iden- only recognizes small insertions and deletions (3, 4). tified von Hippel-Lindau (VHL) tumor suppressor-binding protein 1 Recent evidence demonstrates that eukaryotes contained a sep- (VBP1) as an interacting protein partner for human MSH4 (hMSH4).
    [Show full text]
  • 9Faaa2444c3ffa7b739bc49e981
    Original Article Comparison of Protamine 1 to Protamine 2 mRNA Ratio and YBX2 gene mRNA Content in Testicular Tissue of Fertile and Azoospermic Men Sahar Moghbelinejad, Ph.D.1, 2, Reza Najafipour, Ph.D.1, 2*, Amir Samimi Hashjin, M.Sc.1 1. Cellular and Molecular Research Centre, Qazvin University of Medical Sciences, Qazvin, Iran 2. Department of Medical Genetics, Qazvin University of Medical Sciences, Qazvin, Iran Abstract Background: Although aberrant protamine (PRM) ratios have been observed in infertile men, the mechanisms that implicit the uncoupling of PRM1 and PRM2 expression remain unclear. To uncover these mechanisms, in this observational study we have compared the PRM1/PRM2 mRNA ratio and mRNA contents of two regulatory factors of these genes. Materials and Methods: In this experimental study, sampling was performed by a mul- ti-step method from 50 non-obstructive azoospermic and 12 normal men. After RNA extraction and cDNA synthesis, real-time quantitative polymerase chain reaction (RT- QPCR) was used to analyze the PRM1, PRM2, Y box binding protein 2 (YBX2) and JmjC-containing histone demethylase 2a (JHDM2A) genes in testicular biopsies of the studied samples. Results: The PRM1/PRM2 mRNA ratio differed significantly among studied groups, namely 0.21 ± 0.13 in azoospermic samples and -0.8 ± 0.22 in fertile samples. The amount of PRM2 mRNA, significantly reduced in azoospermic patients. Azoospermic men exhib- ited significant under expression of YBX2 gene compared to controls (P<0.001). mRNA content of this gene showed a positive correlation with PRM mRNA ratio (R=0.6, P=0.007). JHDM2A gene expression ratio did not show any significant difference between the studied groups (P=0.3).
    [Show full text]
  • Genetics of Azoospermia
    International Journal of Molecular Sciences Review Genetics of Azoospermia Francesca Cioppi , Viktoria Rosta and Csilla Krausz * Department of Biochemical, Experimental and Clinical Sciences “Mario Serio”, University of Florence, 50139 Florence, Italy; francesca.cioppi@unifi.it (F.C.); viktoria.rosta@unifi.it (V.R.) * Correspondence: csilla.krausz@unifi.it Abstract: Azoospermia affects 1% of men, and it can be due to: (i) hypothalamic-pituitary dysfunction, (ii) primary quantitative spermatogenic disturbances, (iii) urogenital duct obstruction. Known genetic factors contribute to all these categories, and genetic testing is part of the routine diagnostic workup of azoospermic men. The diagnostic yield of genetic tests in azoospermia is different in the different etiological categories, with the highest in Congenital Bilateral Absence of Vas Deferens (90%) and the lowest in Non-Obstructive Azoospermia (NOA) due to primary testicular failure (~30%). Whole- Exome Sequencing allowed the discovery of an increasing number of monogenic defects of NOA with a current list of 38 candidate genes. These genes are of potential clinical relevance for future gene panel-based screening. We classified these genes according to the associated-testicular histology underlying the NOA phenotype. The validation and the discovery of novel NOA genes will radically improve patient management. Interestingly, approximately 37% of candidate genes are shared in human male and female gonadal failure, implying that genetic counselling should be extended also to female family members of NOA patients. Keywords: azoospermia; infertility; genetics; exome; NGS; NOA; Klinefelter syndrome; Y chromosome microdeletions; CBAVD; congenital hypogonadotropic hypogonadism Citation: Cioppi, F.; Rosta, V.; Krausz, C. Genetics of Azoospermia. 1. Introduction Int. J. Mol. Sci.
    [Show full text]
  • Proteomic Characterization of Transcription and Splicing Factors Associated with a Metastatic Phenotype in Colorectal Cancer
    Proteomic characterization of transcription and splicing factors associated with a metastatic phenotype in colorectal cancer Sofía Torres1+, Irene García-Palmero1+, Consuelo Marín-Vicente1,2, Rubén A. Bartolomé1, Eva Calviño1, María Jesús Fernández-Aceñero3 and J. Ignacio Casal1* +. Equal authorship 1. Functional proteomics. Centro de Investigaciones Biológicas (CIB-CSIC). Ramiro de Maeztu 9. Madrid. Spain. 2. Proteomic facilities. CIB-CSIC. Madrid. Spain 3. Department of Pathology. Hospital Clínico. Madrid. Spain Running title: Transcription factors in metastatic colorectal cancer Keywords: SRSF3, transcription factors, splicing factors, metastasis, colorectal cancer *. Corresponding author J. Ignacio Casal Department of Cellular and Molecular Medicine Centro de Investigaciones Biológicas (CIB-CSIC) Ramiro de Maeztu, 9 28040 Madrid, Spain Phone: +34 918373112 Fax: +34 91 5360432 Email: [email protected] 1 ABSTRACT We investigated new transcription and splicing factors associated with the metastatic phenotype in colorectal cancer. A concatenated tandem array of consensus transcription factors (TFs)-response elements was used to pull down nuclear extracts in two different pairs of colorectal cancer cells, KM12SM/KM12C and SW620/480, genetically-related but differing in metastatic ability. Proteins were analyzed by label-free LC-MS and quantified with MaxLFQ. We found 240 proteins showing a significant dysregulation in highly- metastatic KM12SM cells relative to non-metastatic KM12C cells and 257 proteins in metastatic SW620 versus SW480. In both cell lines there were similar alterations in genuine TFs and components of the splicing machinery like UPF1, TCF7L2/TCF-4, YBX1 or SRSF3. However, a significant number of alterations were cell-line specific. Functional silencing of MAFG, TFE3, TCF7L2/TCF-4 and SRSF3 in KM12 cells caused alterations in adhesion, survival, proliferation, migration and liver homing, supporting their role in metastasis.
    [Show full text]
  • Crossing and Zipping: Molecular Duties of the ZMM Proteins in Meiosis Alexandra Pyatnitskaya, Valérie Borde, Arnaud De Muyt
    Crossing and zipping: molecular duties of the ZMM proteins in meiosis Alexandra Pyatnitskaya, Valérie Borde, Arnaud de Muyt To cite this version: Alexandra Pyatnitskaya, Valérie Borde, Arnaud de Muyt. Crossing and zipping: molecular duties of the ZMM proteins in meiosis. Chromosoma, Springer Verlag, 2019, 10.1007/s00412-019-00714-8. hal-02413016 HAL Id: hal-02413016 https://hal.archives-ouvertes.fr/hal-02413016 Submitted on 16 Dec 2019 HAL is a multi-disciplinary open access L’archive ouverte pluridisciplinaire HAL, est archive for the deposit and dissemination of sci- destinée au dépôt et à la diffusion de documents entific research documents, whether they are pub- scientifiques de niveau recherche, publiés ou non, lished or not. The documents may come from émanant des établissements d’enseignement et de teaching and research institutions in France or recherche français ou étrangers, des laboratoires abroad, or from public or private research centers. publics ou privés. Manuscript Click here to access/download;Manuscript;review ZMM Chromosoma2019_Revised#2.docx Click here to view linked References Crossing and zipping: molecular duties of the ZMM proteins in meiosis 1 2 3 1,2 1,2,* 1,2,* 4 Alexandra Pyatnitskaya , Valérie Borde and Arnaud De Muyt 5 1 Institut Curie, PSL Research University, CNRS, UMR3244, Paris, France. 6 7 2 Paris Sorbonne Université, Paris, France. 8 9 *Valérie Borde, [email protected]; Arnaud De Muyt, [email protected] 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 Keywords : meiosis, crossover, recombination, synaptonemal complex, ZMM 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 1 65 Abstract 1 2 Accurate segregation of homologous chromosomes during meiosis depends on the ability 3 4 of meiotic cells to promote reciprocal exchanges between parental DNA strands, known 5 as crossovers (COs).
    [Show full text]
  • RNA Binding Protein, Ybx2, Regulates RNA Stability During Cold-Induced
    Page 1 of 51 Diabetes RNA binding protein, Ybx2, regulates RNA stability during cold-induced brown fat activation Dan Xu1,2*, Shaohai Xu3, Aung Maung Maung Kyaw2, Yen Ching Lim1, Sook Yoong Chia2, Diana Teh Chee Siang2, Juan R. Alvarez-Dominguez5, Peng Chen3, Melvin Khee-Shing Leow6,7,8, Lei Sun2,4* 1School of Laboratory Medicine and Life Science, Wenzhou Medical University, Wenzhou, Zhejiang 325035, China 2Cardiovascular and Metabolic Disorders Program, Duke-NUS Medical School, 8 College Road, Singapore 169857, Singapore 3Division of Bioengineering, Nanyang Technological University, 70 Nanyang Drive, Singapore 637457, Singapore 4Institute of Molecular and Cell Biology, 61 Biopolis Drive, Proteos, Singapore 138673, Singapore 5Department of Stem Cell and Regenerative Biology, Harvard Stem Cell Institute, Harvard University, 7 Divinity Avenue, Cambridge, MA 02138, USA 6Clinical Nutrition Research Centre, Singapore Institute for Clinical Sciences, Agency for Science, Technology and Research (A*STAR), Singapore, Republic of Singapore. 7Department of Endocrinology, Tan Tock Seng Hospital, 11 Jalan Tan Tock Seng, Singapore 308433, Singapore 8Office of Clinical Sciences, Duke-NUS Medical School, 8 College Road, Singapore 169857, Singapore *Correspondence: [email protected] (D.X.); [email protected] (L.S.) Diabetes Publish Ahead of Print, published online September 29, 2017 Diabetes Page 2 of 51 Abstract Recent years have seen an upsurge of interest on brown adipose tissue (BAT) to combat the epidemic of obesity and diabetes. How its development and activation are regulated at the post-transcriptional level, however, has yet to be fully understood. RNA binding proteins (RBPs) lie in the center of post-transcriptional regulation. To systemically study the role of RBPs in BAT, we profiled >400 RBPs in different adipose depots and identified Y-box binding protein 2 (Ybx2) as a novel regulator in BAT activation.
    [Show full text]
  • Coding and Noncoding Variants in HFM1, MLH3, MSH4, MSH5, RNF212, and RNF212B Affect Recombination Rate in Cattle
    Downloaded from genome.cshlp.org on September 29, 2021 - Published by Cold Spring Harbor Laboratory Press Research Coding and noncoding variants in HFM1, MLH3, MSH4, MSH5, RNF212, and RNF212B affect recombination rate in cattle Naveen Kumar Kadri,1 Chad Harland,1,2 Pierre Faux,1 Nadine Cambisano,1,3 Latifa Karim,1,3 Wouter Coppieters,1,3 Sébastien Fritz,4,5 Erik Mullaart,6 Denis Baurain,7 Didier Boichard,5 Richard Spelman,2 Carole Charlier,1 Michel Georges,1 and Tom Druet1 1Unit of Animal Genomics, GIGA-R & Faculty of Veterinary Medicine, University of Liège (B34), 4000 Liège, Belgium; 2Livestock Improvement Corporation, Newstead, 3240 Hamilton, New Zealand; 3Genomics Platform, GIGA, University of Liège (B34), 4000 Liège, Belgium; 4Allice, 75012 Paris, France; 5GABI, INRA, AgroParisTech, Université Paris-Saclay, 78350 Jouy-en-Josas, France; 6CRV BV, 6800 AL Arnhem, the Netherlands; 7InBioS-Eukaryotic Phylogenomics, Department of Life Sciences and PhytoSYSTEMS, University of Liège (B22), 4000 Liège, Belgium We herein study genetic recombination in three cattle populations from France, New Zealand, and the Netherlands. We identify 2,395,177 crossover (CO) events in 94,516 male gametes, and 579,996 CO events in 25,332 female gametes. The average number of COs was found to be larger in males (23.3) than in females (21.4). The heritability of global recombi- nation rate (GRR) was estimated at 0.13 in males and 0.08 in females, with a genetic correlation of 0.66 indicating that shared variants are influencing GRR in both sexes. A genome-wide association study identified seven quantitative trait loci (QTL) for GRR.
    [Show full text]